Microorganisms attach to surfaces and develop biofilms. Biofilm-associated cells can be differentiated from their suspended counterparts by generation of an extracellular polymeric substance (EPS) matrix, reduced growth rates, and the up- and down- regulation of specific genes. Attachment is a complex process regulated by diverse characteristics of the growth medium, substratum, and cell surface. An established biofilm structure comprises microbial cells and EPS, has a defined architecture, and provides an optimal environment for the exchange of genetic material between cells. Cells may also communicate via quorum sensing, which may in turn affect biofilm processes such as detachment. Biofilms have great importance for public health because of their role in certain infectious diseases and importance in a variety of device-related infections. A greater understanding of biofilm processes should lead to novel, effective control strategies for biofilm control and a resulting improvement in patient management.

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Biofilms: microbial life on surfaces.

Many microorganisms, especially bacteria, produce biosurfactants when grown on water-immiscible substrates. Biosurfactants are more effective, selective, environmentally friendly, and stable than many synthetic surfactants. Most common biosurfactants are glycolipids in which carbohydrates are attached to a long-chain aliphatic acid, while others, like lipopeptides, lipoproteins, and heteropolysaccharides, are more complex. Rapid and reliable methods for screening and selection of biosurfactant-producing microorganisms and evaluation of their activity have been developed. Genes involved in rhamnolipid synthesis (rhlAB) and regulation (rhlI and rhlR) in Pseudomonas aeruginosa are characterized, and expression of rhlAB in heterologous hosts is discussed. Genes for surfactin production (sfp, srfA, and comA) in Bacillus spp. are also characterized. Fermentative production of biosurfactants depends primarily on the microbial strain, source of carbon and nitrogen, pH, temperature, and concentration of oxygen and metal ions. Addition of water-immiscible substrates to media and nitrogen and iron limitations in the media result in an overproduction of some biosurfactants. Other important advances are the use of water-soluble substrates and agroindustrial wastes for production, development of continuous recovery processes, and production through biotransformation. Commercialization of biosurfactants in the cosmetic, food, health care, pulp- and paper-processing, coal, ceramic, and metal industries has been proposed. However, the most promising applications are cleaning of oil-contaminated tankers, oil spill management, transportation of heavy crude oil, enhanced oil recovery, recovery of crude oil from sludge, and bioremediation of sites contaminated with hydrocarbons, heavy metals, and other pollutants. Perspectives for future research and applications are also discussed.

Microbial production of surfactants and their commercial potential.

INTRODUCTION 149 TAXONOMY 149 Historical Features 149 Current Taxonomic Status 149 Delineation of Species 149 Species of Clinical Importance 150 LABORATORY IDENTIFICATION 150 Isolation from Clinical Specimens 150 Morphological, Cultural, and Metabolic Characteristics 151 Species Identification 151 NOSOCOMIAL INFECTIONS CAUSED BY ACINETOBACTER SPP. 152 Overview 152 Respiratory Infection 152 Bacteremia 153 Meningitis 153 Urinary Tract Infection 154 Other Miscellaneous Infections 154 PATHOGENESIS OF ACINETOBACTER INFECTIONS 154 Predisposing Factors 154 Virulence of Acinetobacter spp. 154 EPIDEMIOLOGY 155 Human Carriage 155 Persistence in the Hospital Environment 155 TYPING SYSTEMS 156 Biotyping 156 Antibiograms 156 Serotyping 156 Phage Typing 157 Bacteriocin Typing 157 Protein Profiles 157 Multilocus Enzyme Electrophoretic Typing 157 Plasmid Profiles 157 Analysis by Pulsed-Field Gel Electrophoresis 157 Ribotyping 157 PCR-Based Methods 158 CLINICAL ANTIBIOTIC RESISTANCE 158 BIOCHEMICAL AND GENETIC MECHANISMS OF ANTIBIOTIC RESISTANCE 158 Genetics of Resistance 158 b-Lactams 159 Aminoglycosides 159 Quinolones 160 Other Antibiotics 160 THERAPY OF ACINETOBACTER INFECTIONS 160 CONCLUSIONS 160 ACKNOWLEDGMENTS 161 REFERENCES 161

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Acinetobacter spp. as nosocomial pathogens: microbiological, clinical, and epidemiological features.

Surfactants are surface-active compounds capable of reducing surface and interfacial tension at the interfaces between liquids, solids and gases, thereby allowing them to mix or disperse readily as emulsions in water or other liquids. The enormous market demand for surfactants is currently met by numerous synthetic, mainly petroleum-based, chemical surfactants. These compounds are usually toxic to the environment and non-biodegradable. They may bio-accumulate and their production, processes and by-products can be environmentally hazardous. Tightening environmental regulations and increasing awareness for the need to protect the ecosystem have effectively resulted in an increasing interest in biosurfactants as possible alternatives to chemical surfactants. Biosurfactants are amphiphilic compounds of microbial origin with considerable potential in commercial applications within various industries. They have advantages over their chemical counterparts in biodegradability and effectiveness at extreme temperature or pH and in having lower toxicity. Biosurfactants are beginning to acquire a status as potential performance-effective molecules in various fields. At present biosurfactants are mainly used in studies on enhanced oil recovery and hydrocarbon bioremediation. The solubilization and emulsification of toxic chemicals by biosurfactants have also been reported. Biosurfactants also have potential applications in agriculture, cosmetics, pharmaceuticals, detergents, personal care products, food processing, textile manufacturing, laundry supplies, metal treatment and processing, pulp and paper processing and paint industries. Their uses and potential commercial applications in these fields are reviewed.

Potential commercial applications of microbial surfactants.

Research and development of antimicrobial materials for food applications such as packaging and other food contact surfaces is expected to grow in the next decade with the advent of new polymer materials and antimicrobials. This article reviews the different types of antimicrobial polymers developed for food contact, commercial applications, testing methods, regulations and future trends. Special emphasis will be on the advantages/disadvantages of each technology.

Review of antimicrobial food packaging

Polycationic polymers have been noted for their effects in promoting cell adhesion to various surfaces, but previous studies have failed to describe a mechanism dealing with this type of adhesion. In the present study, three polycationic polymers (chitosan, poly-L-lysine, and lysozyme) were tested for their effects on microbial hydrophobicity, as determined by adhesion to hydrocarbon and polystyrene. Test strains (Escherichia coli, Candida albicans, and a nonhydrophobic mutant, MR-481, derived from Acinetobacter calcoaceticus RAG-1) were vortexed with hexadecane in the presence of the various polycations, and the extent of adhesion was measured turbidimetrically. Adhesion of all three test strains rose from near zero values to over 90% in the presence of low concentrations of chitosan (125 to 250 micrograms/ml). Adhesion occurred by adsorption of chitosan directly to the cell surface, since E. coli cells preincubated in the presence of the polymer were highly adherent, whereas hexadecane droplets pretreated with chitosan were subsequently unable to bind untreated cells. Inorganic cations (Na+, Mg2+) inhibited the chitosan-mediated adhesion of E. coli to hexadecane, presumably by interfering with the electrostatic interactions responsible for adsorption of the polymer to the bacterial surface. Chitosan similarly promoted E. coli adhesion to polystyrene at concentrations slightly higher than those which mediated adhesion to hexadecane. Poly-L-lysine also promoted microbial adhesion to hexadecane, although at concentrations somewhat higher than those observed for chitosan. In order to study the effect of the cationic protein lysozyme, adhesion was studied at 0 degree C (to prevent enzymatic activity), using n-octane as the test hydrocarbon. Adhesion of E. coli increased by 70% in the presence of 80 micrograms of lysozyme per ml. When the negatively charged carboxylate residues on the E. coli cell surface were substituted for positively charged ammonium groups, the resulting cells became highly hydrophobic, even in the absence of polycations. The observed "hydrophobicity" of the microbial cells in the presence of polycations is thus probably due to a loss of surface electronegativity. The data suggest that enhancement of hydrophobicity by polycationic polymers is a general phenomenon.

Mechanism of enhancement of microbial cell hydrophobicity by cationic polymers.

Bad breath is a common phenomenon, usually the result of bacterial metabolism in the oral cavity. It is generally accepted that Gram-negative bacteria are responsible for this problem, largely through degradation of proteinaceous substances. In initial experiments, screening of malodorous isolates following outgrowth of samples obtained from saliva, periodontal pockets, and the tongue dorsum yielded enterobacterial isolates. Clinical studies were conducted to examine the prevalence of such bacteria in four different populations: orthodontic patients, malodor clinic patients, complete-denture wearers, and a healthy young population. The prevalence of Enterobacteriaceae in the oral cavities of the denture-wearing population was very high (48.0%) as compared with the other groups: 27.1% in the malodor clinic patients, 16.4% in the normal population, and 13% among orthodontic patients. Isolates of Klebsiella and Enterobacter emitted foul odors in vitro which resembled bad breath, with concomitant production of volatile sulfides and cadaverine, both compounds related to bad breath. When incubated on a sterile denture, enterobacterial isolates produced typical denture foul odor. Isolates exhibited cell-surface hydrophobic properties when tested for adhesion to acryl and aggregation with ammonium sulphate. The results, taken together, suggest that Klebsiella and related Enterobacteriaceae may play a role in denture malodor.

Isolation of Enterobacteriaceae from the Mouth and Potential Association with Malodor

Bad Breath—Research Perspectives

The ability of Acinetobacter calcoaceticus RAG-1 to adhere to human epithelial cells was investigated and compared with its ability to adhere to a test hydrocarbon (hexadecane). RAG-1, a microorganism originally isolated for growth on hydrocarbon, adhered to epithelial cells when grown under conditions which promote its adherence to hexadecane; similarly, RAG-1 cells adhered poorly to epithelial cells when grown under conditions which cause the cells to possess low affinity towards hexadecane. A mutant derived from RAG-1, MR-481, deficient in its ability to adhere to hydrocarbon, was similarly unable to adhere to epithelial cells. RAG-1 adherence to epithelial cells was not blocked by a number of sugars tested. Streptococcus pyogenes, whose adherence to epithelial cells has been previously attributed to hydrophobic interactions, was also able to adhere to hexadecane. Results suggest that hydrophobic interactions mediate adherence of the strains studied to both epithelial cells and hydrocarbon.

https://iai.asm.org/content/iai/33/1/29.full.pdf

Adherence of Acinetobacter calcoaceticus RAG-1 to human epithelial cells and to hexadecane.

Few scientific investigations have addressed the ability of mouthrinses to reduce oral malodor for periods longer than 3 hours. In the present report, we have employed simple, recently described techniques to assess the day-long reduction in oral malodor of a novel 2-phase oil:water mouthrinse (TPM), as compared to a corresponding placebo rinse, and to a commercial 0.2% chlorhexidine mouthrinse. Sixty dental students were divided randomly into 3 groups, and instructed to use one of the rinses prior to bedtime and the following morning. Measurements carried out in the late afternoon, about 8 to 10 hours following rinsing, were compared with baseline measurements carried out in the late afternoon of the previous day. Volatile sulphide levels were measured using a portable industrial sulphide monitor. Microbial levels were estimated using a simple rinsing technique employing sterilized milk. These quantitative techniques were corroborated by organoleptic (hedonic) ratings of a single odor judge. Both TPM and...

Mel Rosenberg

Papers

Mechanism of enhancement of microbial cell hydrophobicity by cationic polymers.

Isolation of Enterobacteriaceae from the Mouth and Potential Association with Malodor

Bad Breath—Research Perspectives

Adherence of Acinetobacter calcoaceticus RAG-1 to human epithelial cells and to hexadecane.

Day-long reduction of oral malodor by a two-phase oil:water mouthrinse as compared to chlorhexidine and placebo rinses.